Cancer Letters

Cancer Letters

Volume 130, Issues 1–2, 14 August 1998, Pages 93-101
Cancer Letters

A study on p16, pRb, cdk4 and cyclinD1 expression in non-small cell lung cancers

https://doi.org/10.1016/S0304-3835(98)00115-3Get rights and content

Abstract

To observe the expression of p16, pRb, cdk4 and cyclinD1 in non-small cell lung cancers, 104 cases of resected lung cancers were collected, which included squamous cell carcinomas, adenocarcinomas and large cell carcinomas. Immunohistochemistry assay was carried out. The results showed that 67% of squamous cell carcinomas and 46% of adenocarcinomas expressed p16, 64% of squamous cell carcinomas and 85% of adenocarcinomas expressed pRb and 66% of cancers expressed p16 or pRb. About 70% of the tumors expressed cyclinD1. More than 90% of the tumors expressed cdk4 and there was an increased trend with decreasing differentiation of both squamous cell carcinomas and adenocarcinomas. Sixty-seven percent of the highly differentiated and 100% of the poorly differentiated squamous cell carcinomas expressed cdk4. The aberrant p16 and pRb gene product expression played a significant role in the development and histological subtype of lung cancers by conditioning the biological behavior of NSCLC. cdk4 was an important factor in histological differentiation.

Introduction

Non-small cell lung cancer is a malignant tumor which seriously threatens human health. Modern tumor molecular biological studies 1, 2, 3prove that the mechanisms for controlling tumor growth might involve direct regulation of the cell cycle which is regulated by a series of catalytic protein kinase complexes [4]consisting of cyclins (regulatory subunit), cyclin-dependent kinases (cdks) (catalytic subunit) and some others. The activation of corresponding substrates triggers of the corresponding cell phases. cdk4 and cyclinD1 are strongly implicated in the control of G1 progression and the phosphorylation of the retinoblastoma protein (pRb) 5, 6. p16 was found to be altered in many human malignant tumor cell lines and primary tumors 5, 7and p16 protein could specifically interact with cyclinD1 to inhibit the activity of cyclin-dependent kinase4 (cdk4); the latter could catalyze the pRb.

One hundred four cases of resected non-small cell lung cancer (NSCLC) were collected. The expression of p16, pRb, cyclinD1 and cdk4 in NSCLC was investigated by immunohistochemistry (LSAB method). The relationship of expression of p16, pRb, cdk4 and cyclinD1 was studied.

Section snippets

Materials and methods

One hundred four cases of resected NSCLC were collected from our hospital; these included squamous cell carcinomas (n=58), adenocarcinomas (n=41) and large cell carcinomas (n=5). All of the samples were surgically resected and had not previously received either chemotherapy or radiotherapy.

Rabbit polyclonal p16, pRb and cdk4 antibodies were purchased from Santa Cruz Biotechnology and mouse monoclonal cyclinD1 antibody was from NOVO. The DAKO avidin–biotin complex kit was used for detection

Immunostaining with anti-p16 antibody

The nuclei of the positive cells were stained a brownish yellow color (Fig. 1Fig. 2). The p16 expression of tumors was significantly lower than that of normal mucosal epithelium (P<0.001), which implied that the p16 gene was downregulated in tumor cells. The positive expression rate in squamous cell carcinoma and adenocarcinoma was 67.2 and 46%, respectively (Table 1). Additionally, the positive rate of squamous cell tumor was significantly higher than that of adenocarcinoma (χ2=4.32, P<0.05).

Discussion

The absence of p16 expression is a common molecular abnormality in NSCLC 2, 9, 10. In this study, about 40% of lung cancers did not express p16, 52% of cancers partly expressed p16 and 90% of normal mucosal epithelium expressed p16 (P<0.01), which shows that p16 is downregulated in NSCLC and implies that p16 is an important factor in the development of lung cancers. One of the mechanisms of p16 expression was hyperthylation of a G:C rich region within exon 1 of the p16 gene [11]. The expression

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